title: In vitro antibacterial and hemolytic activities of crotamine, a small basic myotoxin from rattlesnake Crotalus durissus.
abstract: Crotamine, a myotoxin from the venom of South American rattlesnake, is structurally related to β-defensins, antimicrobial peptides (AMPs) found in vertebrate animals. Here, we tested the antibacterial properties of crotamine and found that it killed several strains of Escherichia coli, with the MICs ranging from 25 to 100 μg ml⁻¹. Time-kill and bacterial membrane permeabilization assays revealed that killing of bacteria by crotamine occurred within 1 h and reached the maximum by 2 h. Additionally, the anti-E. coli activity of crotamine was completely abolished with 12.5 mM NaCl. Furthermore, the three intramolecular disulfide bonds of crotamine appeared dispensable for its antibacterial activity. The reduced form of crotamine was active against E. coli as well. However, crotamine showed no or weak activity up to 200 μg ml⁻¹ against other species of Gram-negative and Gram-positive bacteria. Crotamine showed no appreciable hemolytic activity to erythrocytes. Our studies revealed that crotamine is also an AMP that kills bacteria through membrane permeabilization. However, crotamine appears to have a narrow antibacterial spectrum, distinct from many classical β-defensins, reinforcing the notion that crotamine originated from the β-defensin gene lineage, but has undergone significant functional diversification.
authors.affiliation: Department of Animal Science, Oklahoma State University, Stillwater, OK, USA. nancyoguiura@butantan.gov.br
authors.forename: Nancy
authors.initials: N
authors.lastname: Oguiura
authors.forename: Malvina
authors.initials: M
authors.lastname: Boni-Mitake
authors.forename: Regina
authors.initials: R
authors.lastname: Affonso
authors.forename: Guolong
authors.initials: G
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology*; D003435:Crotalid Venoms / Q000302:isolation & purification / Q000494:pharmacology*; D017839:Crotalus; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects*; D006090:Gram-Negative Bacteria / Q000187:drug effects*; D006094:Gram-Positive Bacteria / Q000187:drug effects*; D006461:Hemolysis / Q000187:drug effects; D008566:Membranes / Q000378:metabolism; D051379:Mice; D008826:Microbial Sensitivity Tests; D010539:Permeability; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D003435:Crotalid Venoms; C010563:crotamine